Recombinant Human HGF (NS0-expressed) GMP Protein, CF

Catalog #: 294-GMP Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
294-GMP-025

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Recombinant Human HGF (NS0-expressed) GMP Protein Bioactivity

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Product Details

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human HGF (NS0-expressed) GMP Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE with silver staining, under reducing conditions

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to induce IL-11 secretion by Saos‑2 human osteosarcoma cells. Hjertner, O. et al. (1999) Blood 94:3883. The ED₅₀ for this effect is equal or less than 4.00 ng/mL.
The specific activity of recombinant human HGF is >1.0 x 10⁶ IU/mg, which is calibrated against the human HGF WHO International Standard (NIBSC code: 96/564).

Source

Mouse myeloma cell line, NS0-derived human HGF protein
Gln32-Ser728 (Asp384Asn, Asp416Asn, Leu622Ser, His645Arg, Val678Ile)
Manufactured and tested under cGMP guidelines.

Accession #

P14210.2

N-terminal Sequence
Analysis

Amino acid sequencing of the alpha chain was blocked, suggesting it is consistent with Gln₃₂
Gln₃₂-Arg-Lys-Arg-Arg-Asn-Thr-Ile-His-Glu ( alpha -chain)
Val₄₉₅-Val-Asn-Gly-Ile-Pro-Thr-Arg-Thr-Asn ( beta -chain)

Structure / Form

Disulfide-linked heterodimer

Predicted Molecular Mass

alpha chain: 53.7 kDa beta chain: 26 kDa

SDS-PAGE

71-85 kDa, non-reducing conditions
56-65 kDa, and 30-40 kDa, reducing conditions

Product Datasheets

294-GMP

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

294-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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Recombinant Human HGF GMP induces IL-11 secretion by Saos‑2 human osteosarcoma cells. The ED₅₀ for this effect is equal or less than 4.00 ng/mL.

SDS-PAGE

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1 μg/lane of GMP-grade Recombinant Human HGF (Catalog # 294-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing R bands at 30-40 and 56-65 kDa and NR bands at 71-85 kDa, respectively.

Reconstitution Calculator

Background: HGF

HGF, also known as scatter factor and hepatopoietin A, is a pleiotropic protein in the plasminogen subfamily of S1 peptidases. It is a multidomain molecule that includes an N-terminal PAN/APPLE‑like domain, four Kringle domains, and a serine proteinase-like domain that has no detectable protease activity (1‑5). Human HGF is secreted as an inactive 728 amino acid (aa) single chain propeptide. It is cleaved after the fourth Kringle domain by a serine protease to form bioactive
disulfide‑linked HGF with a 60 kDa alpha and 30 kDa beta chain. Alternate splicing generates human HGF isoforms that lack the proteinase‑like domain and different numbers of the Kringle domains. Human HGF shares 91%‑94% aa sequence identity with bovine, canine, feline, mouse, and rat HGF. HGF binds heparan-sulfate proteoglycans and the widely expressed receptor tyrosine kinase, HGF R/c-MET (6, 7). HGF-dependent c-MET activation is implicated in the development of many human cancers (8). HGF regulates epithelial morphogenesis by inducing cell scattering and branching tubulogenesis (9, 10). HGF induces the up‑regulation of integrin alpha 2 beta 1 in epithelial cells by a selective increase in alpha 2 gene transcription (11). This integrin serves as a collagen I receptor, and its blockade disrupts epithelial cell branching tubulogenesis (11, 12). HGF can also alter epithelium morphology by the induction of nectin‑1 alpha ectodomain shedding, an adhesion protein component of adherens junctions (13). In the thyroid, HGF induces the proliferation, motility, and loss of differentiation markers of thyrocytes and inhibits TSH‑stimulated iodine uptake (14). HGF promotes the motility of cardiac stem cells in damaged myocardium (15).

References

Karihaloo, A. et al. (2005) Nephron Exp. Nephrol. 100:e40.
Hammond, D.E. et al. (2004) Curr. Top. Microbiol. Immunol. 286:21.
Rosario, M. and W. Birchmeier (2004) Dev. Cell 7:3.
Lesk, A.M. and W.D. Fordham (1996) J. Mol. Biol. 258:501.
Nakamura, T. et al. (1989) Nature 342:440.
Mizuno, K., et al. (1994) J. Biol. Chem. 269:1131.
Gheradi, E. et al. (2003) Proc. Natl. Acad. Sci. 100:12039.
Corso, S. et al. (2005) Trends Mol. Med. 11:284.
Maeshima, A. et al. (2000) Kid. Int. 58:1511.
Montesano, R. et al. (1991) Cell 67:901.
Chiu, S-J. et al. (2002) J. Biomed. Sci. 9:261.
Saelman, E.U.M. et al. (1995) J. Cell Sci. 108:3531.
Tanaka, Y. et al. (2002) Biochem. Biophys. Res. Commun. 299:472.
Mineo, R. et al. (1994) Endocrinology 145:4355.
Urbanek, K. et al. (2005) Circ. Res. 97:663.

Long Name

Hepatocyte Growth Factor

Entrez Gene IDs

3082 (Human); 15234 (Mouse); 24446 (Rat); 403441 (Canine); 102133907 (Cynomolgus Monkey); 493705 (Feline)

Alternate Names

deafness, autosomal recessive 39; DFNB39; EC 3.4.21; EC 3.4.21.7; fibroblast-derived tumor cytotoxic factor; F-TCF; hepatocyte growth factor (hepapoietin A; scatter factor); Hepatopoeitin-A; Hepatopoietin A; HGF; HGFB; HPTA; HPTAhepatocyte growth factor; lung fibroblast-derived mitogen; Scatter factor; SF; SFhepatopoeitin-A

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.